AmpFLSTR™ Identifiler™ PCR Amplification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Isolate DNA using phenol/chloroform, or magnetic beads (the Maxwell 16 DNA Purification kits, Promega), and MagAttract DNA Mini M48 kit, Qiagen)	Match STR profiles with the ATCC's and German Collection of Microorganisms and Cell cultures' (Braunschweig, Germany) online databases

Upstream tips
Isolate DNA using phenol/chloroform, or magnetic beads (the Maxwell 16 DNA Purification kits, Promega), and MagAttract DNA Mini M48 kit, Qiagen)
Protocol tips
Match STR profiles with the ATCC's and German Collection of Microorganisms and Cell cultures' (Braunschweig, Germany) online databases

Publication protocol

Colon cancer cell lines: Twenty-seven colon cancer cell lines were initially included in the present study. HCT-116, HCT-15, LoVo, RKO, SW1116, SW48, SW620, SW948, NCI-H508 and WiDr were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). ALA, CaCo-2, CO-115, COLO 320, DLD-1, EB, FRI, HT-29, IS1, IS2, IS3, LS1034, LS-174T, TC7, TC71, SW480 and V9P were kindly provided by collaborators. Cell lines were cultured in medium with added fetal bovine serum, glutamine, penicillin and streptomycin, and were maintained in humidified 37 °C 5% CO2 incubators as described in Supplementary Table 2. Before collection, cultures were tested for mycoplasma infection using Myco Alert (Lonza, Walkersville, MD, USA) according to the manufacturer's protocol. DNA was isolated using either a standard phenol/chloroform procedure, or a magnetic beads approach (the Maxwell 16 DNA Purification kits, Promega Corporation, Madison, WI, USA, and MagAttract DNA Mini M48 kit, Qiagen Inc., Valencia, CA, USA). DNA was STR profiled using the AmpFLSTR Identifiler PCR Amplification Kit (Life Technologies, Carlsbad, CA, USA). Resulting cancer cell line STR profiles were cross-compared and, where available, matched with the ATCC's and German Collection of Microorganisms and Cell cultures' (Braunschweig, Germany) online databases. Hierarchical clustering of STR data was performed using Euclidian distances and average linkage clustering in Partek Genomics Suite 6.6 (Partek Inc., St Louis, MO, USA; Figure 2). ALA, CO-115, EB, FRI, IS1, IS2, IS3, TC7, TC71 and V9P are non-commercial cell lines and their STR profiles will be provided upon request. Three of the 27 cancer cell lines were found to be misclassified. ALA and IS2 had identical profiles to SW480/SW620 and LS1034, respectively. TC7 had a STR profile incompatible with its origin as a Caco-2 subclone.41 Consequently, ALA, IS2 and TC7 were excluded from further analysis.

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Manufacturer protocol

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